Skip to main content
Book cover

Liposomes pp 435–444Cite as

Anionic pH Sensitive Lipoplexes

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 605))

Abstract

To provide long circulating nanoparticles which can carry a gene to tumors, we have designed anionic pegylated lipoplexes that are pH sensitive. Anionic pegylated lipoplexes have been prepared from the combined formulation of cationic lipoplexes and pegylated anionic liposomes. The neutralization of the particle surface charge as a function of the pH was monitored by light scattering, in order to determine the ratio between anionic and cationic lipids that would give pH sensitive complexes. This ratio has been optimized to form particles sensitive to pH change in the range 5.5–6.5. Compaction of DNA into these newly formed anionic complexes was checked by DNA accessibility to picogreen. The transfection efficiency and pH sensitive property of these formulations were shown in vitro using bafilomycin, a vacuolar H+-ATPase inhibitor.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Miller A (1998) Cationic liposomes for gene therapy. Angew Chem 41:1768–1785

    Google Scholar 

  2. Nicolazzi C, Garinot M, Mignet N, Scherman D, Bessodes M (2003) Cationic lipids for transfection. Curr Med Chem 10:1263–1277

    Article  CAS  PubMed  Google Scholar 

  3. Eliyahu H, Barenholz Y, Domb A (2005) Polymers for DNA delivery. Molecules 10:34–64

    Article  CAS  PubMed  Google Scholar 

  4. Hirko A, Tang F, Hughes J (2003) Cationic lipid vectors for plasmid DNA delivery. Curr Med Chem 10:1185–1193

    Article  CAS  PubMed  Google Scholar 

  5. Byk G, Wetzer B, Frederic M, Dubertret C, Pitard B, Jaslin G, Scherman D (2000) Reduction-sensitive lipopolyamines as a novel nonviral gene delivery system for modulated release of DNA with improved transgene expression. J Med Chem 43:4377–4387

    Article  CAS  PubMed  Google Scholar 

  6. Luten J, Van Nostrum C, De Smedt S, Hennink W (2008) Biodegradable polymers as non-viral carriers for plasmid DNA delivery. J Control Release 126:97–110

    Article  CAS  PubMed  Google Scholar 

  7. Garinot M, Masson C, Mignet N, Bessodes M, Scherman D (2007) Synthesis and advantages of acid-labile formulations for lipoplexes. In: Gregoriadis G (ed) Liposome technology, vol 1, 3rd edn. CRC Press, Boca Raton, pp 139–163

    Google Scholar 

  8. Martin B, Sainlos M, Aissaoui A, Oudrhiri N, Hauchecorne M, Vigneron JP, Lehn JM, Lehn P (2005) The design of cationic lipids for gene delivery. Curr Pharm Des 3:375–394

    Article  Google Scholar 

  9. Roux E, Stomp R, Giasson S, Pezolet M, Moreau P, Leroux JC (2002) Steric stabilization of liposomes by pH-responsive N-isopropylacrylamide copolymer. J Pharm Sci 91:1795–1802

    Article  CAS  PubMed  Google Scholar 

  10. Kim Y, Park J, Lee M, Kim Y, Park T, Kim S (2005) Polyethylenimine with acid-labile linkages as a biodegradable gene carrier. J Control Release 103:209–219

    Article  CAS  PubMed  Google Scholar 

  11. Shi G, Guo W, Stephenson S, Lee R (2002) Efficient intracellular drug and gene delivery using folate receptor-targeted pH-sensitive liposomes composed of cationic/anionic lipid combinations. J Control Release 80:309–319

    Article  CAS  PubMed  Google Scholar 

  12. Guo X, Szoka FC (2003) Chemical approaches to triggerable lipid vesicles for drug and gene delivery. Acc Chem Res 36:335–341

    Article  CAS  PubMed  Google Scholar 

  13. Lee R, Huang L (1996) Folate-targeted, anionic liposome-entrapped polylysine-condensed DNA for tumor cell-specific gene transfer. J Biol Chem 271:8481–8487

    Article  CAS  PubMed  Google Scholar 

  14. Hafez I, Cullis P (2000) Cholesteryl hemisuccinate exhibits pH sensitive polymorphic phase behavior. Biochim Biophys Acta 1463:107–114

    Article  CAS  PubMed  Google Scholar 

  15. Koynova R, Wang L, Tarahovsky Y, MacDonald R (2005) Lipid phase control of DNA delivery. Bioconjug Chem 16:1335–1339

    Article  CAS  PubMed  Google Scholar 

  16. Hafez I, Ansell S, Cullis P (2000) Tunable pH-sensitive liposomes composed of mixtures of cationic and anionic lipids. Biophys J 79:1438–1446

    Article  CAS  PubMed  Google Scholar 

  17. Simões S, Slepushkin V, Gaspar R, de Lima M, Düzgüneş N (1998) Gene delivery by negatively charged ternary complexes of DNA, cationic liposomes and transferrin or fusigenic peptides. Gene Ther 7:955–964

    Article  Google Scholar 

  18. Zalipsky S, Brandeis E, Newman MS, Woodle MC (1994) Long circulating, cationic liposomes containing amino-PEG-phosphatidylethanolamine. FEBS Lett 353:71–74

    Article  CAS  PubMed  Google Scholar 

  19. Masson C, Garinot M, Mignet N, Wetzer B, Mailhe P, Scherman D, Bessodes M (2004) pH sensitive PEG lipids containing orthoester linkers: new potential tools for nonviral gene delivery. J Control Release 99:423–434

    Article  CAS  PubMed  Google Scholar 

  20. Choi J, MacKay J, Szoka F (2003) Low-pH-sensitive PEG-stabilized plasmid-lipid nanoparticles: preparation and characterization. Bioconjugate Chem 14:420

    Article  CAS  Google Scholar 

  21. Kale A, Torchilin V (2007) Enhanced transfection of tumor cells in vivo using “Smart” pH-sensitive TAT-modified pegylated liposomes. J Drug Target 15:537–548

    Article  Google Scholar 

  22. Mignet N, Richard C, Seguin J, Largeau C, Bessodes M, Scherman D (2008) Anionic pH-sensitive pegylated lipoplexes to deliver DNA to the tumors. Int J Pharm 361(1–2):194–201

    Article  CAS  PubMed  Google Scholar 

  23. Thompson B, Mignet N, Hofland H, Lamons D, Seguin J, Nicolazzi C, de la Figuera N, Kuen R, Meng Y, Scherman D, Bessodes M (2005) Neutral post-grafted colloidal particles for gene delivery. Bioconjug Chem 16:608–614

    Article  CAS  PubMed  Google Scholar 

  24. Mignet N, Cadet M, Bessodes M, Scherman D (2007) Incorporation of PEG lipid into lipoplexes: on-line incorporation assessment, and pharmacokinetics advantages. In: Gregoriadis G (ed) Liposome technology, vol 2, 3rd edn. CRC Press, Boca Raton, pp 273–292

    Google Scholar 

  25. Tranchant I, Thompson B, Nicolazzi C, Mignet N, Scherman D (2004) Physicochemical optimization of plasmid delivery by cationic lipids. J Gene Med 1(Suppl 1):S24–S35

    Article  Google Scholar 

Download references

Acknowledgments

The author would like to thank Caroline Richard for her dedicated work during her fellowship and Michel Bessodes for providing the anionic cholesterol derivatives.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Nathalie Mignet .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Mignet, N., Scherman, D. (2010). Anionic pH Sensitive Lipoplexes. In: Weissig, V. (eds) Liposomes. Methods in Molecular Biology, vol 605. Humana Press. https://doi.org/10.1007/978-1-60327-360-2_30

Download citation

  • DOI: https://doi.org/10.1007/978-1-60327-360-2_30

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60327-359-6

  • Online ISBN: 978-1-60327-360-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics